High-speed fiber scanning endoscope for volumetric multi-megahertz optical coherence tomography.

We present a forward-viewing fiber scanning endoscope (FSE) for high-speed volumetric optical coherence tomography (OCT). The reduction in size of the probe was achieved by substituting the focusing optics by an all-fiber-based imaging system which consists of a combination of scanning single-mode fibers, a glass spacer, made from a step-index multi-mode fiber, and a gradient-index fiber. A lateral resolution of 11 µm was achieved at a working distance of 1.2 mm. The newly designed piezo-based FSE has an outer diameter of 1.6 mm and a rigid length of 13.5 mm. By moving the whole imaging optic in spirals for scanning the sample, the beam quality remains constant over the entire field of view with a diameter of 0.8 mm. The scanning frequency was adjusted to 1.22 kHz for use with a 3.28 MHz Fourier domain mode locked OCT system. Densely sampled volumes have been imaged at a rate of 6 volumes per second.

Thermo-elastic optical coherence tomography.

The absorption of nanosecond laser pulses induces rapid thermo-elastic deformation in tissue. A sub-micrometer scale displacement occurs within a few microseconds after the pulse arrival. In this Letter, we investigate the laser-induced thermo-elastic deformation using a 1.5 MHz phase-sensitive optical coherence tomography (OCT) system. A displacement image can be reconstructed, which enables a new modality of phase-sensitive OCT, called thermo-elastic OCT. An analysis of the results shows that the optical absorption is a dominating factor for the displacement. Thermo-elastic OCT is capable of visualizing inclusions that do not appear on the structural OCT image, providing additional tissue type information.

High-resolution rectoscopy using MHz optical coherence tomography: a step towards real time 3D endoscopy.

Colonoscopy and endoscopic ultrasound play pivotal roles in the assessment of rectal diseases, especially rectal cancer and inflammatory bowel diseases. Optical coherence tomography (OCT) offers a superior depth resolution, which is a critical factor for individualizing the therapeutic concept and evaluating the therapy response. We developed two distinct rectoscope prototypes, which were integrated into a 1300 nm MHz-OCT system constructed at our facility. The rapid rotation of the distal scanning probe at 40,000 revolutions per minute facilitates a 667 Hz OCT frame rate, enabling real-time endoscopic imaging of large areas. The performance of these OCT-rectoscopes was assessed in an ex vivo porcine colon and a post mortem human in-situ colon. The OCT-rectoscope consistently distinguished various layers of the intestinal wall, identified gut-associated lymphatic tissue, and visualized a rectal polyp during the imaging procedure with 3D-reconstruction in real time. Subsequent histological examination confirmed these findings. The body donor was preserved using an ethanol-glycerol-lysoformin-based technique for true-to-life tissue consistency. We could demonstrate that the novel MHZ-OCT-rectoscope effectively discriminates rectal wall layers and crucial tissue characteristics in a post mortem human colon in-situ. This real-time-3D-OCT holds promise as a valuable future diagnostic tool for assessing disease state and therapy response on-site in rectal diseases.

Microscope-integrated optical coherence tomography for in vivo human brain tumor detection with artificial intelligence.

OBJECTIVE: It has been shown that optical coherence tomography (OCT) can identify brain tumor tissue and potentially be used for intraoperative margin diagnostics. However, there is limited evidence on its use in human in vivo settings, particularly in terms of its applicability and accuracy of residual brain tumor detection (RTD). For this reason, a microscope-integrated OCT system was examined to determine in vivo feasibility of RTD after resection with automated scan analysis. METHODS: Healthy and diseased brain was 3D scanned at the resection edge in 18 brain tumor patients and investigated for its informative value in regard to intraoperative tissue classification. Biopsies were taken at these locations and labeled by a neuropathologist for further analysis as ground truth. Optical OCT properties were obtained, compared, and used for separation with machine learning. In addition, two artificial intelligence-assisted methods were utilized for scan classification, and all approaches were examined for RTD accuracy and compared to standard techniques. RESULTS: In vivo OCT tissue scanning was feasible and easily integrable into the surgical workflow. Measured backscattered light signal intensity, signal attenuation, and signal homogeneity were significantly distinctive in the comparison of scanned white matter to increasing levels of scanned tumor infiltration (p < 0.001) and achieved high values of accuracy (85%) for the detection of diseased brain in the tumor margin with support vector machine separation. A neuronal network approach achieved 82% accuracy and an autoencoder approach 85% accuracy in the detection of diseased brain in the tumor margin. Differentiating cortical gray matter from tumor tissue was not technically feasible in vivo. CONCLUSIONS: In vivo OCT scanning of the human brain has been shown to contain significant value for intraoperative RTD, supporting what has previously been discussed for ex vivo OCT brain tumor scanning, with the perspective of complementing current intraoperative methods for this purpose, especially when deciding to withdraw from further resection toward the end of the surgery.

The neurosurgical benefit of contactless in vivo optical coherence tomography regarding residual tumor detection: A clinical study.

Purpose: In brain tumor surgery, it is crucial to achieve complete tumor resection while conserving adjacent noncancerous brain tissue. Several groups have demonstrated that optical coherence tomography (OCT) has the potential of identifying tumorous brain tissue. However, there is little evidence on human in vivo application of this technology, especially regarding applicability and accuracy of residual tumor detection (RTD). In this study, we execute a systematic analysis of a microscope integrated OCT-system for this purpose. Experimental design: Multiple 3-dimensional in vivo OCT-scans were taken at protocol-defined sites at the resection edge in 21 brain tumor patients. The system was evaluated for its intraoperative applicability. Tissue biopsies were obtained at these locations, labeled by a neuropathologist and used as ground truth for further analysis. OCT-scans were visually assessed with a qualitative classifier, optical OCT-properties were obtained and two artificial intelligence (AI)-assisted methods were used for automated scan classification. All approaches were investigated for accuracy of RTD and compared to common techniques. Results: Visual OCT-scan classification correlated well with histopathological findings. Classification with measured OCT image-properties achieved a balanced accuracy of 85%. A neuronal network approach for scan feature recognition achieved 82% and an auto-encoder approach 85% balanced accuracy. Overall applicability showed need for improvement. Conclusion: Contactless in vivo OCT scanning has shown to achieve high values of accuracy for RTD, supporting what has well been described for ex vivo OCT brain tumor scanning, complementing current intraoperative techniques and even exceeding them in accuracy, while not yet in applicability.

Continuous spectral zooming for in vivo live 4D-OCT with MHz A-scan rates and long coherence.

We present continuous three-dimensional spectral zooming in live 4D-OCT using a home-built FDML based OCT system with 3.28 MHz A-scan rate. Improved coherence characteristics of the FDML laser allow for imaging ranges up to 10 cm. For the axial spectral zoom feature, we switch between high resolution and long imaging range by adjusting the sweep range of our laser. We present a new imaging setup allowing for synchronized adjustments of the imaging range and lateral field of view during live OCT imaging. For this, a novel inline recalibration algorithm was implemented that enables numerical k-linearization of the raw OCT fringes for every frame instead of every volume. This is realized by acquiring recalibration data within the dead time of the raster scan at the turning points of the fast axis scanner. We demonstrate in vivo OCT images of fingers and hands at different resolution modes and show real three-dimensional zooming during live 4D-OCT. A three-dimensional spectral zooming feature for live 4D-OCT is expected to be a useful tool for a wide range of biomedical, scientific and research applications, especially in OCT guided surgery.

Registration of histological brain images onto optical coherence tomography images based on shape information.

Identifying tumour infiltration zones during tumour resection in order to excise as much tumour tissue as possible without damaging healthy brain tissue is still a major challenge in neurosurgery. The detection of tumour infiltrated regions so far requires histological analysis of biopsies taken from at expected tumour boundaries. The gold standard for histological analysis is the staining of thin cut specimen and the evaluation by a neuropathologist. This work presents a way to transfer the histological evaluation of a neuropathologist onto optical coherence tomography (OCT) images. OCT is a method suitable for real timein vivoimaging during neurosurgery however the images require processing for the tumour detection. The method demonstrated here enables the creation of a dataset which will be used for supervised learning in order to provide a better visualization of tumour infiltrated areas for the neurosurgeon. The created dataset contains labelled OCT images from two different OCT-systems (wavelength of 930 nm and 1300 nm). OCT images corresponding to the stained histological images were determined by shaping the sample, a controlled cutting process and a rigid transformation process between the OCT volumes based on their topological information. The histological labels were transferred onto the corresponding OCT images through a non-rigid transformation based on shape context features retrieved from the sample outline in the histological image and the OCT image. The accuracy of the registration was determined to be 200 ± 120µm. The resulting dataset consists of 1248 labelled OCT images for each of the two OCT systems.

Differentiation of different stages of brain tumor infiltration using optical coherence tomography: Comparison of two systems and histology.

The discrimination of tumor-infiltrated tissue from non-tumorous brain tissue during neurosurgical tumor excision is a major challenge in neurosurgery. It is critical to achieve full tumor removal since it directly correlates with the survival rate of the patient. Optical coherence tomography (OCT) might be an additional imaging method in the field of neurosurgery that enables the classification of different levels of tumor infiltration and non-tumorous tissue. This work investigated two OCT systems with different imaging wavelengths (930 nm/1310 nm) and different resolutions (axial (air): 4.9 µm/16 µm, lateral: 5.2 µm/22 µm) in their ability to identify different levels of tumor infiltration based on freshly excised ex vivo brain samples. A convolutional neural network was used for the classification. For both systems, the neural network could achieve classification accuracies above 91% for discriminating between healthy white matter and highly tumor infiltrated white matter (tumor infiltration >60%) .This work shows that both OCT systems with different optical properties achieve similar results regarding the identification of different stages of brain tumor infiltration.

Cavity length control for Fourier domain mode locked (FDML) lasers with µm precision.

In highly dispersion compensated Fourier domain mode locked (FDML) lasers, an ultra-low noise operation can only be achieved by extremely precise and stable matching of the filter tuning period and light circulation time in the cavity. We present a robust and high precision closed-loop control algorithm and an actively cavity length controlled FDML laser. The cavity length control achieves a stability of ∼0.18 mHz at a sweep repetition rate of ∼418 kHz which corresponds to a ratio of 4×10-10. Furthermore, we prove that the rapid change of the cavity length has no negative impact on the quality of optical coherence tomography using the FDML laser as light source.

Flexible A-scan rate MHz-OCT: efficient computational downscaling by coherent averaging.

In order to realize adjustable A-scan rates of fast optical coherence tomography (OCT) systems, we investigate averaging of OCT image data acquired with a MHz-OCT system based on a Fourier Domain Mode Locked (FDML) laser. Increased system sensitivity and image quality can be achieved with the same system at the cost of lower imaging speed. Effectively, the A-scan rate can be reduced in software by a freely selectable factor. We demonstrate a detailed technical layout of the strategies necessary to achieve efficient coherent averaging. Since there are many new challenges specific to coherent averaging in swept source MHz-OCT, we analyze them point by point and describe the appropriate solutions. We prove that coherent averaging is possible at MHz OCT-speed without special interferometer designs or digital phase stabilization. We find, that in our system up to ∼100x coherent averaging is possible while achieving a sensitivity increase close to the ideal values. This corresponds to a speed reduction from 3.3 MHz to 33 kHz and a sensitivity gain of 20 dB. We show an imaging comparison between coherent and magnitude averaging of a human finger knuckle joint in vivo with 121 dB sensitivity for the coherent case. Further, the benefits of computational downscaling in low sensitivity MHz-OCT systems are analyzed.

Correction: Live video rate volumetric OCT imaging of the retina with multi-MHz A-scan rates.

[This corrects the article DOI: 10.1371/journal.pone.0213144.].

Live video rate volumetric OCT imaging of the retina with multi-MHz A-scan rates.

Surgical microscopes are vital tools for ophthalmic surgeons. The recent development of an integrated OCT system for the first time allows to look at tissue features below the surface. Hence, these systems can drastically improve the quality and reduce the risk of surgical interventions. However, current commercial OCT-enhanced ophthalmic surgical microscopes provide only one additional cross sectional view to the standard microscope image and feature a low update rate. To present volumetric data at a high update rate, much faster OCT systems than the ones applied in today's surgical microscopes need to be developed. We demonstrate live volumetric retinal OCT imaging, which may provide a sufficiently large volume size (330x330x595 Voxel) and high update frequency (24.2 Hz) such that the surgeon may even purely rely on the OCT for certain surgical maneuvers. It represents a major technological step towards the possible application of OCT-only surgical microscopes in the future which would be much more compact thus enabling many additional minimal invasive applications. We show that multi-MHz A-scan rates are essential for such a device. Additionally, advanced phase-based OCT techniques require 3D OCT volumes to be detected with a stable optical phase. These techniques can provide additional functional information of the retina. Up to now, classical OCT was to slow for this, so our system can pave the way to holographic OCT with a traditional confocal flying spot approach. For the first time, we present point scanning volumetric OCT imaging of the posterior eye with up to 191.2 Hz volume rate. We show that this volume rate is high enough to enable a sufficiently stable optical phase to a level, where remaining phase errors can be corrected. Applying advanced post processing concepts for numerical refocusing or computational adaptive optics should be possible in future with such a system.

Ultra low noise Fourier domain mode locked laser for high quality megahertz optical coherence tomography.

We investigate the origin of high frequency noise in Fourier domain mode locked (FDML) lasers and present an extremely well dispersion compensated setup which virtually eliminates intensity noise and dramatically improves coherence properties. We show optical coherence tomography (OCT) imaging at 3.2 MHz A-scan rate and demonstrate the positive impact of the described improvements on the image quality. Especially in highly scattering samples, at specular reflections and for strong signals at large depth, the noise in optical coherence tomography images is significantly reduced. We also describe a simple model that suggests a passive physical stabilizing mechanism that leads to an automatic compensation of remaining cavity dispersion in FDML lasers.

Combined 60° Wide-Field Choroidal Thickness Maps and High-Definition En Face Vasculature Visualization Using Swept-Source Megahertz OCT at 1050 nm.

PURPOSE: To demonstrate ultrahigh-speed swept-source optical coherence tomography (SS-OCT) at 1.68 million A-scans/s for choroidal imaging in normal and diseased eyes over a ∼60° field of view. To investigate and correlate wide-field three-dimensional (3D) choroidal thickness (ChT) and vascular patterns using ChT maps and coregistered high-definition en face images extracted from a single densely sampled Megahertz-OCT (MHz-OCT) dataset. METHODS: High-definition, ∼60° wide-field 3D datasets consisting of 2088 × 1024 A-scans were acquired using a 1.68 MHz prototype SS-OCT system at 1050 nm based on a Fourier-domain mode-locked laser. Nine subjects (nine eyes) with various chorioretinal diseases or without ocular pathology are presented. Coregistered ChT maps, choroidal summation maps, and depth-resolved en face images referenced to either the retinal pigment epithelium or the choroidal-scleral interface were generated using manual segmentation. RESULTS: Wide-field ChT maps showed a large inter- and intraindividual variance in peripheral and central ChT. In only four of the nine eyes, the location with the largest ChT was coincident with the fovea. The anatomy of the large lumen vessels of the outer choroid seems to play a major role in determining the global ChT pattern. Focal ChT changes with large thickness gradients were observed in some eyes. CONCLUSIONS: Different ChT and vascular patterns could be visualized over ∼60° in patients for the first time using OCT. Due to focal ChT changes, a high density of thickness measurements may be favorable. High-definition depth-resolved en face images are complementary to cross sections and thickness maps and enhance the interpretation of different ChT patterns.

High definition live 3D-OCT in vivo: design and evaluation of a 4D OCT engine with 1 GVoxel/s.

We present a 1300 nm OCT system for volumetric real-time live OCT acquisition and visualization at 1 billion volume elements per second. All technological challenges and problems associated with such high scanning speed are discussed in detail as well as the solutions. In one configuration, the system acquires, processes and visualizes 26 volumes per second where each volume consists of 320 x 320 depth scans and each depth scan has 400 usable pixels. This is the fastest real-time OCT to date in terms of voxel rate. A 51 Hz volume rate is realized with half the frame number. In both configurations the speed can be sustained indefinitely. The OCT system uses a 1310 nm Fourier domain mode locked (FDML) laser operated at 3.2 MHz sweep rate. Data acquisition is performed with two dedicated digitizer cards, each running at 2.5 GS/s, hosted in a single desktop computer. Live real-time data processing and visualization are realized with custom developed software on an NVidia GTX 690 dual graphics processing unit (GPU) card. To evaluate potential future applications of such a system, we present volumetric videos captured at 26 and 51 Hz of planktonic crustaceans and skin.